A polyclonal goat antiserum against the calcium-binding protein calretininis a versatile tool for various immunochemical techniques

Specific antibodies are useful tools to label particular neurons and at tim es to delineate neuronal circuits - a task not easily achieved by other tec hniques. Human recombinant calretinin, a protein belonging to the EF-hand f amily of Ca2+-binding proteins, was used to produce an antiserum in goat. T he specificity of the antiserum to recognize calretinin was demonstrated in brain extracts from mouse, rat, and chick and in extracts from human tumor cell lines known to express this protein. Immunohistochemically, the antis erum-stained specific neurons in human, rhesus monkey, mouse, and rat brain . The goat anti-calretinin antiserum is an appropriate tool for double- or triple-immunolabeling studies along with previously-established rabbit and mouse antibodies, Thus, it allows for the concomitant staining with antibod ies directed against other EF-hand calcium-binding proteins including calbi ndin-D-28k and parvalbumin. The antiserum can further be used for the quant ification of calretinin in different tissues or cell lines in a sandwich EL ISA. Additionally, it is well suited for the detection of calretinin in cer tain cell lines or malignant pleural mesotheliomas. Immunostaining of these samples is comparable to that with the well-characterized calretinin-speci fic polyclonal rabbit antiserum 7696. (C) 1999 Elsevier Science B.V. All ri ghts reserved.